Protein redistribution in TCR-directed NF-kB activation

抗原刺激的T细胞分裂和效应子功能的获得依赖于信号传导。 由T细胞受体（TCR）指导的转导，以及随后的许多免疫调节因子的激活， 影响基因表达变化的转录因子。NF-KB家族的转录因子有 在这个过程中至关重要。将研究TCR调节的NF-κ B活化途径 通过分子生物化学和高度先进的成像技术的结合。目标是 建立1）蛋白质-蛋白质相互作用，2）亚细胞蛋白质 组织和3）TCR指导的NF-κ B中细胞质信号转导物的生物化学修饰 通路这一目标将通过执行三个具体目标来实现： 目标1.确定信号传导的局部蛋白质-蛋白质关联的机制意义 TCR介导的NF-κ B活化的中间体。我们将检验TCR介导的NF-κ B 信号传导涉及到特定蛋白质复合物的有序组装和分解， 亚细胞结构域，与特定的蛋白质复合物发挥关键的机械作用。 目标二。在单个细胞中，研究抗原信号 信号转导复合物的强度、组装和TCR指导的NF-κ B的成功活化 信号转导途径实验的目的是为了评估以下假设， 通过TCR的抗原信号被转换成无活化或完全活化的“二元”结果 效应器功能。此外，这些功能结果依赖于信号转导决定 在单细胞水平上发生的事件。 目标3。为了明确磷酸化和MALT 1相互作用结构域在TCR中的作用， 介导的BcHO降解和激活。我们将进行实验来检验这一假设， BcHO的激活和蛋白水解破坏是机械耦合的，并且这两个过程都是 通过BcHO与MALT 1的相互作用以及BcHO的PKC依赖性磷酸化来调节。 这些研究将有助于建立基础研究基础， 通过高度特异性抑制抗原受体发挥作用的新型免疫调节药物 介导的NF-κ B信号级联的激活。

Antigen-stimulated T cell division and acquisition of effector functions are dependent upon signal transduction directed by the T cell receptor (TCR), and the consequent activation of a number of transcription factors that effect changes in gene expression. Transcription factors of the NF-KB family are of central importance in this process. The TCR-regulated NF-KB activation pathway will be investigated through a combination of molecular, biochemical, and highly advanced imaging techniques. The goal is to establish mechanistic relationships between 1) protein-protein interactions, 2) subcellular protein organization and 3) biochemical modification of cytoplasmic signal transducers in the TCR-directed NF-KB pathway. This goal will be accomplished via the execution of three specific aims: Aim1. Todetermine the mechanistic significance of localizedprotein-protein associations of signaling intermediates in TCR-directed activation of NF-KB. We will test the hypothesis that TCR-directed NF-KB signaling involves the orchestrated assembly and disassembly of defined protein complexes into discrete subcellular domains, with specific protein complexes playing a critical mechanistic role. Aim 2. Toinvestigate, in individual cells, the mechanistic relationship between antigen signal strength, assembly of signal transduction complexes, and successful activation of the TCR-directed NF-KB signal transduction pathway. Experiments will be directed towards the evaluation of the hypothesis that antigen signals through the TCR are converted into "binary" outcomes of either no activation or full activation of effector functions. Furthermore, these functional outcomes are dependent on signal transduction decision events made at the single-cell level. Aim 3. To define the roles of phosphorylation and the MALT1 interaction domain in the TCR- mediated degradation and activation ofBcHO. We will perform experiments to test the hypothesis that activation and proteolytic destruction of BcHO are mechanistically coupled, and that both processes are regulated by BcHO interaction with MALT1 and by PKC-dependent phosphorylation of BcHO. These studies will help establish the basic research foundation that could lead to the development of novel immuno-modulatory drugs, which would function via highly specfic inhibition of antigen-receptor mediated activation of the NF-KB signaling cascade.